Enzyme engineering is an important part of the modern biotechnology. Industrial biocatalysis is considered the third wave of biotechnology following pharmaceutical and agricultural waves. In 25 years, China has made a mighty advances in enzyme engineering research. This review focuses on enzyme genomics, enzyme proteomics, biosynthesis, microbial conversion and biosensors in the Chinese enzyme engineering symposiums and advances in enzyme preparation industry in China.
Agriculture ; Biocatalysis ; Biosensing Techniques ; Biotechnology ; China ; Enzymes ; Industrial Microbiology ; Protein Engineering ; Proteomics

ObjectiveTo comprehend the change of the characteristics of lifting force produced by a working space marker in process of its cavity-forming. Methods37 patients were successively operated with the surgical mode of minimally invasive video-assisted thyroidectomy (22/37 cases received a lobotomy and others un derwent a partial thyroidectomy) from January to August, 2010. Instead of hand-retraction, a mechanical armworking space marker type I ( WSM-I, MIEO Medinstr Co. Ltd, China) was applied to establish a working space. After pathway making, an interlayer-cavity above the lobe was created by the space maker and endoscopic view was properly built. Following all these steps, a simulated space making procedure was performed in a way of stepwise hook-lifting (5 mm rising per time). The lifting force ( LF)was measured during the process with a modflied force-measure device (FB-50, DESIK company, Germen). Then recorded data were assessed and analyzed statistically. Results①Ascending scope of LF in the process of entire space-forming was 0-27.5 Newton (N).②Along with hook rising, LF ascended correspondingly and 2 specific values emerged: One was 11. 2 ±3.5 N,as the lifting height approached 1.5 cm ( also a approximate position of essential space-forming ( Pe), at which themusculo-cutaneoustissuejustbecame tight) ; the other was 17.5 ± 4.3 N , as the lifting height approached 1.75 cm ( also a approximate position of maximal space-forming (Pmax), at which the musculo-cutaneous tissue appeared real tight, but not in a status of extreme tightness). ③Two types of LF ascending were found when the values transferred to a curve diagram : a palliative linearity increasing while lifting height varied from 0 to 1.5 cm (PO to Pe) and a rapid exponent-like increasing while lifting height varied from 1.5 to 1.75cm ( Pe to Pmax). ④ Dependability analyses yielded a diverse statistical outcome: negative significance of the comparison between incision length and LF value ( P ＞ 0. 05 ), and positive significance of the comparison between skin thickness and LF value ( P ＜ 0. 01 ). Conclusions①LF produced by WSM-I while establishing a working space is proper and relatively small, since the maximal value is merely 27.5N, far less than the stress produced by ordinary cosmetic skin expansion. ②The whole space-forming process can be divided into 2 stages according to the characteristic of LF ascending which correspond also separately to the “essential cavity-forming” and “the maximum cavity-forming” in the real establishing of a working space. ③Attention should be paid to the later stage since in which a rapid LF increasing occurs while the appearance of musculo-cutaneous tissue changes from “just become tight” to “real appear tight”. ④LF control, especially the fine readjustment at or about Pmax should be of necessity in individual space-forming, and then, ideal working space establishment can be archived at a pre cisely balanced LF point: maximum cavity volume acquired and minimal tissue expansion stress produced.

Objective To investigate the stability of lumbar single?level dynamicly fixed by socket track pedicle screws after decompression. Methods Socket track pedicle screws are made of titanium?alloy, swing like universal screw within the scope of 18° in the sagittal plane and 8° in the horizontal plane. Motion constraints were due to the hole which the screw goes through. Six pig spines (L1-5) were tested by applying a pure moment of 4.018 Nm in 3 directions of loading. The range of motion (ROM) of the following groups underwent different surgical procedures was measured:(1) intact;(2) dynamic fixation 1 (decompres?sion by total laminectomy and instrumentation with socket track pedicle screws); (3) dynamic fixation 2 (decompression by total laminectomy and bilateral facetectomy while instrumentation with socket track pedicle screws);( 4) instability resulted from decom?pression (decompression by total laminectomy plus bilateral facetectomy without any rod);(5) hybrid fixation (decompression by to?tal laminectomy and bilateral facetectomy while instrumentation with 2 socket track pedicle screws connected with 2 unilateral pedicle screws ). Results Compared with the intact specimens (in flexion, extension, lateral bending and rotation, corresponding angular displacements were 1.84°±0.75°, 1.55°±0.34°, 2.2°±1.07°, 2.06°±0.76° respectively), the ROM of segment L3, 4 of each group were significantly restricted except for axial rotation. In flexion, extension and lateral bending, the ROM was reduced to 0.71°±0.39°, 0.46°±0.18°, 0.85°±0.66° in dynamic fixation 1 group;0.79°±0.43°, 0.71°±0.20°, 1.17°±0.48° in dynamic fixation 2 group and 0.63°±0.50°, 0.37°±0.19°, 0.54°±0.60° in hybrid fixation group. The rotation ROM of dynamic fixation 1 group and hy?brid fixation group were decreased to 1.50°±0.64° and 1.22°±0.36°, whereas the ROM of dynamic fixation 2 had a non?significant reduction to 1.85°±0.67°. The extension and rotation ROM of dynamic fixation 2 increased significantly comparing to dynamic fixa?tion 1 group; the extension, lateral bending and axial rotation ROM of hybrid fixation group were reduced comparing to the state of dynamic fixation 2 group. Conclusion The dynamic instrumentation of single?level lumbar spine with socket track pedicle screws could offer stability in all directions. The flexion, extension, lateral bending and rotation ROM of spine with dynamic instru?mentation after total laminectomy and bilateral facetectomy could be reduced.

From air bacteria capable of decomposing creatinine, three single independent strains K9510、K9511 and K9512 have been isolated. The highest creatinine amidohydrolase (EC 3.5.2.10; creatininase) producing strain K9510 was screened out. The strain K9510 was identified as Pseudomonas sp. The results of culture condition for creatininase formation by strain K9510 were obtained as follows: creatinine and creatine were found to be the effective inducers for enzyme formation; the solution of mixed metallic salts could stimulate cell growth and enzyme formation. The suitable medium for creatininase formation was consisted of 0.9% creatinine、 0. 15% yeast extract、 0. 09% malt extract、0.05% NH4C1 and some amount of the solution of mixed metallic salts at pH5. 5. When the bacterium was grown in 250mL conic flask containing 50mL of the medium mentioned above on the rotary shaker(250r/min) at 35℃ for 33 h, about 50 u creatininase was obtained.

Objective To evaluate the role of phosphoinositide 3 kinase/protein kinase B (PI3K/Akt) signaling pathway in hydrogen-induced inhibition of cell apoptosis during myocardial ischemia/reperfusion (I/R) in rats.Methods Forty healthy male Sprague-Dawley rats,weighing 300-350 g,were randomly allocated into 4 groups (n =10 each) using a random number table:sham operation group (S group),I/R group,hydrogen group (group H),and hydrogen + LY294002 group (group HL).Myocardial I/R was induced by occlusion of the anterior descending branch of left coronary artery for 30 min followed by 120 min reperfusion.In H and HL groups,99.6 ％ hydrogen 5 ml/kg was injected intraperitoneally immediately after beginning of reperfusion,and in addition LY294002 (0.3 mg/kg) was injected through the caudal vein before hydrogen injection in group HL.Arterial blood samples were collected at the end of 120 min reperfusion for determination of serum creatine kinase isoenzyme-MB (CK-MB) and lactate dehydrogenase (LDH) activities.The rats were then sacrificed.Myocardial apoptosis was detected by TUNEL and apoptosis index (AI) was calculated.The expression of Bcl-2,Bax and caspase-3 was detected by immuno-histochemistry.Bcl-2/Bax ratio was calculated.Results The serum CK-MB and LDH activities and AI were significantly increased,the expression of myocardial Bcl-2,Bax and caspase-3 was upregulated,and the ratio of Bcl-2/Bax was decreased in group I/R as compared with group S.Compared with group I/R,the serum CK-MB and LDH activities and AI were significantly decreased,the expression of myocardial Bcl2 was up-regulated,while the expression of myocardial Bax and caspase-3 was down-regulated,and the ratio of Bcl-2/Bax was increased in group H,and no significant changes were found in the parameters mentioned above in group HL.The serum CK-MB and LDH activities and AI were significantly increased,the expression of myocardial Bcl-2 was down-regulated,while the expression of myocardial Bax and caspase-3 was up-regulated,and Bcl-2/Bax ratio was decreased in group HL as compared with group H.Conclusion Hydrogen can activate the PI3K/Akt signaling pathway,and further up-regulates Bcl-2 expression and down-regulates Bax and Caspase-3 expression,thus inhibiting cell apoptosis during myocardial I/R in rats.

Objective To investigate the effect of inhibition of glycogen synthase kinase?3 beta ( GSK?3β) activity on sevoflurane postconditioning?induced cardioprotection in diabetic rats. Methods Healthy adult male Sprague?Dawley rats, weighing 250-300 g, in which diabetes mellitus was induced by intraperitoneal 1% streptozotocin 60 mg∕kg combined with high?fat and high?sucrose diet and confirmed by blood glucose level >16. 7 mmol∕L. Forty rats with diabetes mellitus were divided into 5 groups ( n=8 each) using a random number table: sham operation group ( S group ) , ischemia?reperfusion ( I∕R ) group, sevoflurane postconditioning group ( SP group) , GSK?3β inhibitor SB216763 group ( SB group) , and sevoflurane postconditioning plus SB216763 group ( SS group ) . Myocardial ischemia was induced by 30 min occlusion of the left anterior descending branch of the coronary artery followed by 120 min reperfu?
sion. The rats inhaled sevoflurane with the end?tidal concentration of 2.5% for 5 min starting from 1 min be?fore reperfusion in group SP. SB216763 0.2 mg∕kg was injected via the caudal vein at 1 min before reperfu?sion in group SB. In group SS, the rats inhaled sevoflurane with the end?tidal concentration of 2.5% for 5 min starting from 1 min before reperfusion, and SB216763 0.2 mg∕kg was injected via the caudal vein at 1 min before reperfusion. At 120 min of reperfusion, blood samples were collected from the carotid artery for determination of serum creatine kinase?MB (CK?MB) activity and cardiac troponin I (cTnI) concentra?tions. Myocardial specimens were collected at 120 min of reperfusion for microscopic examination of the pathological changes and for determination of myocardial infarct size ( by 2,3,5?triphenyltetrazolium chlo?ride staining) and phosphorylated GSK?3β (p?GSK?3β) expression (by Western blot). Results Com?pared with group S, the myocardial infarct size and serum CK?MB activity and cTnI concentration were sig?nificantly increased, and the expression of p?GSK?3βwas significantly down?regulated in I∕R, SP, SB and SS groups (P<0.05). Compared with group I∕R, the myocardial infarct size and serum CK?MB activity and cTnI concentration were significantly decreased, and the expression of p?GSK?3β was significantly up?regulated in SB and SS groups (P<0.05), and no significant change was found in the parameters men?tioned above in group SP ( P>0.05) . Compared with group SB, the myocardial infarct size and serum CK?MB activity and cTnI concentration were significantly decreased, and the expression of p?GSK?3β was sig?nificantly up?regulated in group SS (P<0.05). The pathological changes of myocardium were significantly attenuated in SB and SS groups as compared with group I∕R and group SP . Conclusion Inhibition of GSK?3β activity can improve sevoflurane postconditioning?induced cardioprotection in diabetic rats.

Background and purpose:It has been demonstrated that radical prostatectomy for patients with oligometastatic prostate cancer may contribute to improving local control of prostate cancer and overall survival by several retrospective studies. Perioperative complications play an important role in determining whether radical prostatectomy is appropriate for patients with oligometastatic prostate cancer. This study aimed to discuss the recurrence rate and the sever-ity of perioperative complications, and the primary curative effect of radical prostatectomy on oligometastatic prostate can-cer patients.Methods:A total number of 247 patients who received radical prostatectomy were recruited in the study from Jul. 2015 to Jan. 2016, including 25 patients with oligometastatic prostate cancer and 222 patients with localized prostate cancer. Patients with perioperative complications in both groups were graded with the Clavien-Dindo grading system. The proportion of PSA decline and the rates and severity of perioperative complications were analyzed in both groups.Results:The cases of prostate specific antigen (PSA) decline in the oligometastatic group were 21 (84.0%), lower than the localized group with 212 cases (95.5%). There were 6 cases (24.0%) with postoperative complications in the oligometastatic group, including serious complications (Ⅲ or above) 1 case (4.0%), and 49 cases (22.1%) with postoperative complications in the localized group, including serious complications (Ⅲ or above) 7 cases (3.2%). The differences between the groups reached no statistical significance (P>0.05).Conclusion:Radical prostatectomy for patients with oligometastatic prostate cancer could be safe, effective, and appropriate, the risk of perioperative complications should not be one of the limiting factors.

ObjectiveTo realize the relevance between hypertension and serum uric acid (SUA) in residents in Baise City.MethodsTwenty one thousand,five hundred and eighty eight Baise residents were examined.The body weight,height,waist circumference,hip circumference,blood pressure(BP) were recorded and blood lipidlevels,blood glucoseand serum uric acid (SUA) were tested.The relevance between hypertension and hyperuricemia (HUA) was analyzed by multi-factor variance analysis,x2 test,t test,linear regression and Logistic regression analysis.ResultsHypertension prevalence rate was 31.5％,the frequency of HUA was 12.0％,and 5.8％ residents had both.For patients with both hypertension and HUA,when compared to those with normal serum uric level,their mean diastolic blood pressure(DBP) was(84±10),(72±6) mm Hg res-pectively,there mean systolic blood pressure (SBP) was(151±12),(127±6) mm Hg respectively,and pulse pressure (PP) was (50±12),(37±8) mm Hg respectively.The average BMI of those two groups was (23.9±2.6),(21.7±2.4) kg/m2 respectively.The average TC level was (6.0±1.3),(5.2±1.1)mmol/L respectively,the average TG level was(2.3±2.2),(1.5±0.7) mmol/L respectively,and the average LDL-C level was(3.4±1.3),(3.0±1.1) mmol/L respectively.All these parameters were higher than those people without hypertension or HUA(P＜0.01).However,the situation of HDL-C[ (1.2±0.4),(1.5±0.5)mmol/L] was the opposite (P＜0.01).Regression analysis had shown that age,BMI,waist-hip ratio,SUA,blood glucose,TC,TG,HDL-C and LDL-C were independent risk factors for hypertension,while people with HUA was 3.1 times more when compared to those without HUA.The prevalence rate of HUA in people with hypertension was 2.6 times higher than that of the control group.ConclusionHypertension and HUA are two public health problems that prevalent in the residents in Baise City.Obesity,high blood glucose level and hyperlipidemia are thebasis for the development of hypertension and HUA.Hypertension can be the cause of HUA and verse versa.They may have mutual interactions.The adverse effect of HUA on-cardiovascular system should be taken into consideration clinically.

There were 3 000 Zhuangs,1 102 Miaos, and 1 283 Yaos in Baise City of Guangxi,who were enrolled in this population sampling stratfying survey. Height, weight, blood pressure, fasting blood glucose, and lipids were determined, and compared with those of 2 000 Hans of the same town. The prevalence of hypertension and metablic syndrome in Zhuang inhabitants was high, so were the disorders of glycemia and lipidemia, while in Miao and Yao minorities, the prevalences were comparatively lower. The awareness, treatment, and control of hypertension in these minorities were insufficient.